Citations

Reagents

… C. difficile TcdA and TcdB toxoids (formaldehyde inactivated; List Biological Laboratories Inc. Campbell, USA) were used at 10µg/mL and 5µg/mL respectively. …

Experimental autoimmune encephalomyelitis immunization

C57BL/6 mice were immunized subcutaneously (s.c.) in the flank with 200 μg of MOG35–55 peptide emulsified in complete Freund´s adjuvant (CFA) containing 0.1 mg Mycobacterium tuberculosis (Becton Dickinson) along with an intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories Inc.) on the day of immunization and two days later. …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Experimental design animal study
A schematic representation of the experimental timeline is depicted in Fig. 1B. Mice were orally exposed, by using a blunt needle, to 0.5 mL raw milk, the milk samples heated at 50 °C, 60 °C, 65 °C, 70 °C, 75 °C, 80 °C or PBS (as a control) for eight consecutive days (days-9 to -2). Subsequently, mice (n = 8 per group) were orally (by means of gavage) sensitized to 20 mg ovalbumin (OVA; 20 mg per 0.5 mL PBS; grade V; Sigma-Aldrich) using 15 μg cholera toxin (CT; List Biological Laboratories, Campbell, CA, USA) as an adjuvant (days 0, 7, 14, 21, and 28). …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

… Anthrax lethal factor (#172C) and Anthrax protective antigen (#171E) were from List Biological Labs, Inc. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

Reagents
… Cholera toxin List Biological Laboratories Cat#100B …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Treg cell isolation and adoptive transfer

… Diphtheria toxin (List Biologicals, CA) was administered via intraperitoneal injection in 100µL of sterile PBS in the following doses and days relative to influenza A virus infection (day 0): 50 mcg/Kg on day -2 and 10 mcg/Kg every 48 hours starting on day 0 and ending on day 28 post-infection. Five days later, 1 x 106 young or aged splenic Treg cells in 100 µL of sterile PBS were transferred via retro-orbital injection into the
influenza-treated Foxp3DTR mice.

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Enzyme-linked immunosorbent assay(ELISA)

… To measure the levels of IgG against CCP2, we used a commercially available ELISA
kit (Euro-Diagnostica, Malmö, Sweden). For the detection of anti-inuenza, and anti-diphtheria toxin
antibodies, 96-well plates were coated with 1 μg/ml of A/H1N1 subunit (Bio-Rad, Hercules, CA, USA) and
diphtheria toxin (List Biological Laboratories, Campbell, CA, USA), respectively. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Reagents and cell lines.

… Pertussis toxin was from List Biologicals …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

… Mice also received 200 ng of Bordetella pertussis toxin (List Biological Laboratories) intraperitoneally on days 0 and 2. …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Experiment 3:

… Upon examining the sections through the PBel in animals with injections of AAV-FLEX-ArchT-GFP into the DR of Sert-Cre mice, we found intense innervation of the external lateral part of the parabrachial (PBel) area. To further confirm that PBel receives innervation from serotonergic neurons in the DR, we also injected Sert-L10 mice (n = 4) with retrograde tracer, cholera toxin subunit b (24 nl, 0.2% – CTb, List Biological Laboratories Inc.) unilaterally in the lateral PB. One week later these mice were sacrificed, brains perfused, and tissue sections processed for immunohistochemistry for CTb using the procedure described below.

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt